Novel Strategy Using Tryptic Peptide Immunoaffinity-Based LC–MS/MS to Quantify Denosumab in Monkey Serum

Abstract

Aim: Denosumab is a recombinant fully human IgG2 that has a high affinity and specificity for human RANKL. Commercially available RANKL labeled with an Fc fragment cannot be used to establish an indirect ELISA. To characterize denosumab pharmacokinetic a robust and accuracy method should be developed urgently. Results: In this study, an immunoaffinity enrichment method coupled with LC–MS/MS was established. The LC–MS/MS method acquired a linear range from 0.1 to 30 μg/ml. The intra- and inter-run precision (CV%) was within 11.5 and 10.5%, respectively. More importantly, the LC–MS/MS pharmacokinetic data were consistent with ELISA. Conclusion: This approach accelerated the quantification, reduced the costs and provided an alternative in case of lacking the special antigen to denosumab or a RANKL-biotinylated reagent.

Keywords::

• denosumab
• ELISA
• immunoaffinity enrichment
• LC–MS/MS
• RANKL

Supplementary Data

Financial & competing interests disclosure

The authors are grateful to the award to Zimei Wu from the Taishan Scholar Project, which helped fund this research. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

Acknowledgements

The authors are grateful to the award to Z Wu from the Taishan Scholar Project, which helped fund this research.